H(2)O(2) Damages the Stemness of Rat Bone Marrow-Derived Mesenchymal Stem Cells: Developing a “Stemness Loss” Model

BACKGROUND: The number of patients with spinal cord injury caused by motor vehicle accidents, violent injuries, and other types of trauma increases year by year, and bone marrow mesenchymal stem cell (BMSC) transplants are being widely investigated to treat this condition. However, the success rate of BMSCs transplants is relatively low due to the presence of oxidative stress in the new microenvironment. Our main goals in the present study were to evaluate the damaging effects of H(2)O(2) on BMSCs and to develop a model of “stemness loss” using rat BMSCs. MATERIAL/METHODS: Bone marrow-derived mesenchymal stem cells were obtained from the bone marrow of young rats reared under sterile conditions. The stem cells were used after 2 passages following phenotypic identification. BMSCs were divided into 4 groups to evaluate the damaging effects of H(2)O(2): A. blank control; B. 100 uM H(2)O(2); C. 200 uM H(2)O(2) and D. 300 uM H(2)O(2). The ability of the BMSCs to differentiate into 3 cell lineages and their colony formation and migration capacities were analyzed by gene expression, colony formation, and scratch assays. RESULTS: The cells we obtained complied with international stem cell standards demonstrated by their ability to differentiate into 3 cell lineages. We found that 200–300 uM H(2)O(2) had a significant effect on the biological behavior of BMSCs, including their ability to differentiate into 3 cell lineages, the expression of stemness-related proteins, and their migration and colony formation capacities. CONCLUSIONS: H(2)O(2) can damage the stemness ability of BMSCs at a concentration of 200–300 uM.